TSEG-2基因在小鼠睾丸扭转复位模型中的表达特征

目的 探讨睾丸特异表达基因2(testis specific expressed gene 2,TSEG-2)在小鼠睾丸扭转复位模型中的表达特征.方法 昆明小鼠36只,随机分组为对照组(6只)、假手术组(6只)、单侧睾丸扭转复位实验组(24只).实验组分为2组,每组12只,左侧睾丸扭转720°维持2 h,分别于复位后1、7 d取扭转侧睾丸.采用HE染色、原位末端标记技术(TUNEL)观察睾丸组织形态改变;黄嘌呤氧化酶法、硫代巴比妥酸比色法测定超氧化物歧化酶(SOD)、丙二醛(MDA)活性;原位杂交法观测TSEG-2在睾丸生精细胞内的表达定位;实时定量PCR法检测TSEG-2基因在睾丸组织中的表达水平.结果 对照组和假手术组生精上皮排列规则,扭转复位后1、7 d的睾丸组织内生精上皮结构松散,出现生精细胞凋亡,Johnsen's评分分别降低23.4%、64.1%(P<0.01),SOD活性降低11.6%、22.2%(P<0.05),MDA活性升高69.6%、93.2%(P<0.01).TSEG-2基因表达定位于小鼠睾丸生精小管的精原细胞和精母细胞.与对照组比较,扭转复位1、7 d后睾丸组织内TSEG-2表达水平分别上调2.2倍、6.6倍(P<0.01).结论 成功建立小鼠睾丸扭转复位模型,TSEG-2表达上调可能与抗氧化酶活性下降、生精细胞凋亡有关.

Expression of testis specific expressed gene 2 in murine testicular torsion/detorsion model

Objective To explore the expression profiles of testis specific expressed gene 2 (TSEG-2)in murine testicular torsion/detorsion(TD)model.Methods Thirty six Kunming mice were randomly divided into control(n=6),sham(n=6),and experimental(n=24)groups.In the experimental group,the left testes of the TD mice were rotated 720 degrees for 2 hrs,and then detorsed.The testes were harvested after 1 and 7 days of detorsion.H&E staining and TUNEL staining were performed for morphological and apoptotic studies.The activities of superoxide disnmtase(SOD)and malondialdehycle(MDA)were measured by xanthine superoxidase and thiobarbiuric acid,respectively.The TSEG-2 expression in testes was detected by in situ hybridization and real-time quantitative PCR.Results Compared with the serniniferous tubules of the control and sham operated mice,the sparse structure and apoptotic spermatogenic cells of the seminiferous tubules significantly increased in the mice subjected to TD.One day or 7 days after TD,the Johnsen's score was reduced by 23.4% and 64.1%,respectively(P<0.01).The SOD activities were decreased by 11.6%and 22.2%,respectively(P<0.05).The MDA increased by 69.6%and 93.2%,respectively(P<0.01).The positive staining for TSEG-2 mRNA localized at the spermatogonia and spermatocytes of seminiferous tubules.One day or 7 days after TD,the TSEG-2 expression was enhanced by 2.2 and 6.6 folds,respectively.Conclusions The enhanced TSEG-2 expression may be correlated with impaired anti-oxidative activities and incrassated apoptosis of spermatogenic cells in the murine testicular torsion/detorsion model.